Thiol-mediated Uptake of a Cysteine-containing Nanobody for Anti-Cancer Drug Delivery

Goerdeler, F.; Reuber, E. E.; Luehle, J.; Leichnitz, S.; Freitag, A.; Nedielkov, R.; Moeller, H. M.; Seeberger, P. H.; Moscovitz, O.

doi:10.1101/2022.07.12.497993

bioRxiv · 10.1101/2022.07.12.497993

Thiol-mediated Uptake of a Cysteine-containing Nanobody for Anti-Cancer Drug Delivery

Goerdeler, F.·Reuber, E. E.·Luehle, J.·Leichnitz, S.·Freitag, A.·Nedielkov, R.·Moeller, H. M.·Seeberger, P. H.·Moscovitz, O.

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Abstract

The identification of tumor-specific biomarkers is one of the bottlenecks in the development of cancer therapies. Previous work revealed altered surface levels of reduced/oxidized cysteines in many cancers due to overexpression of redox-controlling proteins such as protein disulfide isomerases on the cell surface. Alterations in surface thiols can promote cell adhesion and metastasis, making thiols attractive targets for treatment. Only a few tools are available to study surface thiols on cancer cells and exploit them for theranostics. Here, we describe a nanobody (CB2) that recognizes B cell lymphoma in a thiol-dependent manner. CB2 binding strictly requires the presence of a non-conserved cysteine in the antigen-binding region and correlates with elevated surface levels of free thiols on B cell lymphoma compared to healthy lymphocytes. Nanobody CB2 can induce complement-dependent cytotoxicity against lymphoma cells when functionalized with synthetic rhamnose trimers. Lymphoma cells internalize CB2 in a thiol-mediated manner such that the nanobody can be used to deliver cytotoxic agents. Hence, surface thiols can be used as lymphoma biomarkers and targeted by thiol-binding nanobodies. Functionalization of internalizable CB2 is the basis for a range of diagnostic and therapeutic applications of this thiol-binding nanobody. TOC Graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=69 SRC="FIGDIR/small/497993v1_ufig1.gif" ALT="Figure 1"> View larger version (20K): org.highwire.dtl.DTLVardef@1f47ba9org.highwire.dtl.DTLVardef@1e2bd41org.highwire.dtl.DTLVardef@f7151eorg.highwire.dtl.DTLVardef@18b7681_HPS_FORMAT_FIGEXP M_FIG C_FIG SynopsisNanobody CB2 specifically binds and internalizes into B cell lymphoma via thiol-based interactions. Functionalized CB2 can be used for complement recruitment or drug delivery to lymphoma cells.

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Goerdeler, F., Reuber, E. E., Luehle, J., Leichnitz, S., Freitag, A., Nedielkov, R., Moeller, H. M., Seeberger, P. H., Moscovitz, O.. 2022-07-14. Thiol-mediated Uptake of a Cysteine-containing Nanobody for Anti-Cancer Drug Delivery. https://doi.org/10.1101/2022.07.12.497993

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